--Eleven tips for operational researchers working with health programmes: our experience based on implementing differentiated tuberculosis care in south India.

Due to the workload and lack of a critical mass of trained operational researchers within their ranks, health systems and programmes may not be able to dedicate sufficient time to conducting operational research (OR). Hence, they may need the technical support of operational researchers from research/academic organisations. Additionally, there is a knowledge gap regarding implementing differentiated tuberculosis (TB) care in programme settings. In this 'how we did it' paper, we share our experience of implementing a differentiated TB care model along with an inbuilt OR component in Tamil Nadu, a southern state in India. This was a health system initiative through a collaboration of the State TB cell with the Indian Council of Medical Research institutes and the World Health Organisation country office in India. The learnings are in the form of eleven tips: four broad principles (OR on priority areas and make it a health system initiative, implement simple and holistic ideas, embed OR within routine programme settings, aim for long-term engagement), four related to strategic planning (big team of investigators, joint leadership, decentralised decision-making, working in advance) and three about implementation planning (conducting pilots, smart use of e-tools and operational research publications at frequent intervals). These may act as a guide for other Indian states, high TB burden countries that want to implement differentiated care, and for operational researchers in providing technical assistance for strengthening implementation and conducting OR in health systems and programmes (TB or other health programmes). Following these tips may increase the chances of i) an enriching engagement, ii) policy/practice change, and iii) sustainable implementation.

Isolation, characterisation, anticancer and anti-oxidant activities of 2-methoxy mucic acid from Rhizophora apiculata: an in vitro and in silico studies.

The main objective of the present study is to isolate and characterise the novel bioactive molecule, 2-methoxy mucic acid (4) from Rhizophora apiculate Blume under the Rhizophoraceae family. In this study, the 2-methoxy mucic acid (4) was isolated for the first time from the methanolic extract of the leaves of R. apiculata. Anticancer activity of 2-methoxy mucic acid (4) was evaluated against HeLa and MDA-MB-231 cancer cell lines and they displayed promising activity with IC50 values of 22.88283 ± 0.72 µg/ml in HeLa and 2.91925 ± 0.52 µg/ml in the case of MDA-MB-231, respectively. Furthermore, the antioxidant property of 2-methoxy mucic acid (4) was found to be (IC50) 21.361 ± 0.41 µg/ml. Apart from in vitro studies, we also performed extensive in silico studies (molecular docking and molecular dynamics simulation) on four critical antiapoptotic Bcl-2 family members (Bcl-2, Bcl-w, Bcl-xL and Bcl-B) towards 2-methoxy mucic acid (4). The results revealed that this molecule showed higher binding affinity towards Bcl-B protein (ΔG = -5.8 kcal/mol) and the structural stability of this protein was significantly improved upon binding of this molecule. The present study affords vital insights into the importance of 2-methoxy mucic acid (4) from R. apiculata. Furthermore, it opens the therapeutic route for the discovery of anticancer drugs. Research HighlightsThis is a first report on a bioactive compound identified and characterised; a novel 2-methoxy mucic acid derived from methanolic crude extract from the leaves of R. apiculata from ANI.Estimated binding free energy of 2-methoxy mucic acid is found to be -5.8 kcal/mol to the anti-apoptotic Bcl-B protein.2-methoxy mucic acid showed both significant anti-cancer and anti-oxidant activity.Communicated by Ramaswamy H. Sarma.

Incidence and effectiveness of manipulation under anaesthesia for stiffness following primary total knee arthroplasty.

Background: There are various modalities of cor- rection of stiffness following total knee arthro-plasty. Manipulation under anaesthesia (MUA) is generally indicated for people who fail to achieve their pre operative range of motion at 12 weeks. The purpose of this study was to determine: (1) the effect of MUA on Flexion arc (2) the influence of timing of MUA from index procedure and of diabetes mellitus on final flexion achieved. Methods: We retrospectively evaluated patients who underwent manipulation following total knee arthroplasty at our institution between January 2016 to December 2018. For the purpose of analysis, we have divided the patients into two groups. Those who underwent manipulation within 12 weeks and later than 12 weeks. We have also compared the effect of MUA between diabetic and non-diabetic patients. All were operated with posterior stabilised (PS) prosthesis by a single senior arthroplasty surgeon. The final flexion achieved during their last clinical follow-up were recorded and compared with the pre MUA flexion. Results: The incidence of MUA after TKA at our institute during this period is about 1.14 %. There was a significant statistical difference between the pre and post manipulation flexion, with p value <0.01. There was no significant statistical difference between those who were manipulated before 12 weeks and after 12 weeks in improving the Flexion of the operated knees. We have found that both the diabetic and non diabetic group had comparable flexion after the manipulation in our study. Conclusion: Manipulation after anaesthesia is a safe first intervention to improve post operative stiffness and gain additional range of motion following TKA in patients who develop stiffness. It can be done even after 12 weeks of surgery with reasonably good gain in range of motion.

A comprehensive in silico and in vitro studies on quinizarin: a promising phytochemical derived from Rhizophora mucronata Lam.

Mangrove plants are a great source of phytomedicines, since from the beginning of human civilization and the origin of traditional medicines. In the present study, ten different mangrove leaf methanolic extracts were screened for the type of phytochemicals followed by assessing antimicrobial, anti-oxidant and anti-cancer activities. The efficient methanolic crude extract of Rhizospora mucornata was further purified and characterized for the presence of the bioactive compound. Based on UV-visible spectroscopy, FTIR, NMR and HRMS analysis, the bioactive compound was 1,4-dihydroanthraquinone; also termed as Quinizarin. This identified compound was potential in exhibiting antimicrobial, antioxidant, and cytotoxic activity. Quinizarin inhibited the growth of Bacillus cereus and Klebsiella aerogenes with minimum inhibitory concentration (MIC) of 0.78 and 1.5 mg/ml. The DPPH free radical scavenging assay revealed the maximum activity of 99.8% at the concentration of 200 µg/ml with an IC50 value of 12.67 ± 0.41 µg/ml. Cytotoxic assay against HeLa (cervical) and MDA-MB231(breast) cancer cell lines revealed IC50 values to be 4.60 ± 0.26 and 3.89 ± 0.15 µg/ml. Together the results of molecular docking and molecular dynamics simulation studies explained that Quinizarin molecule showed stronger binding affinity (-6.2 kcal/mol) and significant structural stability towards anti-apoptotic Bcl-2 protein. Thus, the study put forth the promising role of the natural molecule - Quinizarin isolated from R. mucornata in the formulation of therapeutic drugs against bacterial infections and cancer. Communicated by Ramaswamy H. Sarma.

Biological evaluation of gallic acid and quercetin derived from Ceriops tagal: insights from extensive in vitro and in silico studies.

Gallic acid (PubChem CID: 370) and quercetin (PubChem CID: 5280343) are major phenolic compounds in many mangrove plants that have been related to health cure. In the present study, the active fractions namely gallic acid (1) and quercetin (2) were isolated from the methanolic extract of leaves of Ceriops tagal in a Tropical mangrove ecosystem of Andaman and Nicobar Island (ANI), India. The chemical structures were determined by spectroscopic analysis: Fourier-Transform Infrared spectroscopy (FT-IR), 1H, 13C Nuclear Magnetic Resonance (NMR) spectroscopy, and High-resolution mass spectrometry (HRMS). The anticancer activity of isolated compounds (1) and (2) were evaluated by in vitro assays against two human cancer cell lines namely, HeLa (Cervical) and MDA-MB231 (Breast) cancer cells revealed that IC50 values of gallic acid (HeLa: 4.179197 ± 0.45 µg/ml; MDA-MB231: 80.0427 ± 0.19 µg/ml at 24 h) and quercetin (HeLa: 99.914 ± 0.18 µg/ml; MDA-MB231: 18.288382 ± 0.12 µg/ml at 24 h), respectively. Antioxidant properties of gallic acid (1) and quercetin (2) are found to be IC50 value of 0.77 ± 0.41 µg/ml and 1.897 ± 0.81 µg/ml, respectively. Molecular docking results explained that gallic acid (1) and quercetin (2) showed estimated binding free energy (ΔG) of -5.4 and -6.9 kcal/mol towards drug target Bcl-B protein, respectively. The estimated inhibition constant (Ki) for these two molecules are 110 and 8.75 µM, respectively. The MD simulation additionally supported that quercetin molecule is significantly improved the structural stability of Bcl-B protein. The present study provides key insights about the importance of polyphenols, and thus leads to open the therapeutic route for anti-cancer drug discovery process.Communicated by Ramaswamy H. Sarma.

A Review on Antidiabetic Properties of Indian Mangrove Plants with Reference to Island Ecosystem.

Mangrove ecosystem has many potential species that are traditionally used by the coastal communities for their traditional cure for health ailments as evidenced by their extensive uses to treat hepatic disorders, diabetes, gastrointestinal disorders, anti-inflammation, anticancer, and skin diseases, etc. In recent times, the diabetes mellitus (DM), a serious physiological disorder all over the world, occur due to the relative or complete deficiency of insulin in the body, characterized by an abnormally high blood glucose level. India has a rich traditional knowledge on plant-based drug formulations that are protective and curative for many health ailments. In this context, we aimed to compile the works done on the antidiabetic activities of mangrove species from Indian coastal regions especially on Andaman and Nicobar Islands as well as some recent works reported from other countries. A total of 126 published articles and 31 mangrove species related pieces of information were gathered with reference to antidiabetic properties of mangroves. This review summarizes the chemical structures, molecular formula, molecular weight, and their biological activities with an aspiration that it might be helpful for the future bioprospecting industries who are interested in develop the natural drugs for DM.

Microwave assisted extraction of pectin from waste Citrullus lanatus fruit rinds.

In this present study, microwave-assisted extraction (MAE) was applied to extraction of pectin from waste Citrullus Lanatus fruit rinds. Extraction parameters which are employed in this study are microwave power (160-480 W), irradiation time (60-180s), pH (1-2) and solid-liquid ratio (1:10-1: 30 g/ml) and they were optimized using a four factor three levels Box-Behnken response surface design (BBD) coupled with desirability function methodology. The results showed that, all the process variables have significant effect on the extraction yield of pectin. Optimum MAE conditions for the highest pectin yield from waste C. Lanatus fruit rinds (25.79%) were obtained with microwave power of 477 W, irradiation time of 128 s, pH of 1.52, solid-liquid ratio of 1:20.3g/ml respectively. Validation experiment results were well agreed with predicted value.

Optimization of microwave assisted extraction of pectin from orange peel.

In this study, microwave-assisted extraction was applied for pectin extraction from the dried orange peel and Box-Behnken response surface design was used to study and optimize the effects of processing variables (microwave power, irradiation time, pH and solid-liquid ratio) on the yield of pectin. The amount of pectin extracted increased with increasing microwave power, while it reduces as the time, pH and solid-liquid ratio increased. From the results, second order polynomial model was developed and it adequately explained the data variation and significantly represented the actual relationship between independent variables and the response. An optimization study using Derringer's desired function methodology was performed and optimal conditions based on both individual and combinations of all independent variables (microwave power of 422W, irradiation time of 169 s, pH of 1.4 and solid-liquid ratio of 1:16.9 g/ml) were determined with maximum pectin yield of 19.24%, which was confirmed through validation experiments.

Nanoemulsified ethanolic extract of Pyllanthus amarus Schum & Thonn ameliorates CCl4 induced hepatotoxicity in Wistar rats.

Phyllanthus amarus (PA) is commonly used in traditional medicine for hepatoprotectivity. The major limitation is that, treatment requires a large quantity of herbal extract for a longer duration. Aim of the present study was to encapsulate ethanolic plant extract for sustained release of constituents in intestine and facilitate maximum absorption. The efficacy was compared for the hepatoprotective activity of nanoencapsulated ethanolic extract of P. amarus (NPA) and PA in carbon tetrachloride (CCl4) induced hepatotoxic male rats. Based on total phenol content (TPC), the loading efficiency of nanocapsules was 89% (pH 7.0) and optimum concentration was 2:18 (mg/mL) for plant extract: olive oil. Scanning electron microscopy (SEM) showed a spherical morphology, photon correlation spectroscopy (PCS) identified mean particle diameter as 213 nm and Fourier transform infrared spectroscopy (FT-IR) revealed that the phytoconstituents were stable. An oral dose of NPA (20 mg/kg body wt.) showed a better hepatoprotective activity than PA (100 mg/kg body wt.) and also repeated dose oral toxicity proved to be safe. These biochemical assessments were supported by rat biopsy examinations. In conclusion, the nanoemulsification method may be applied for poor water-soluble ethanolic herbal extracts to reduce the dosage and time.

Aminonaphthoquinones--a novel class of compounds with potent antimalarial activity against Plasmodium falciparum.

Malaria is a major tropical disease, which kills two million people annually. The population at risk from this disease has increased because of the difficulties in eradicating the mosquito vector in the endemic regions and the emergence and spread of parasite resistance to all the commonly used antimalarials. Since antimalarials are the major arsenal for treatment of the disease, there is an urgent need for newer drugs with novel mechanisms of action, which will be effective against all strains of the parasite. As a part of our anti-infective drug discovery program, we have investigated 18 compounds including several synthetic and natural naphthoquinones as potential antimalarial agents. We have identified aminonaphthoquinones, as a class of antimalarial compounds with antimalarial activity against Plasmodium falciparum. Among these compounds, 2-amino-3-chloro-1,4-naphthoquinone is the most potent. It had an IC(50)of 0.18 micro M (37.3 ng ml(-1)) against the W2 clone, and is more potent than chloroquine, which had an IC(50)of 0.23 micro M (72 ng ml(-1)). It was also active against the D6 clone. In general, 2-amino-1,4-naphthoquinone analogs and the 4-amino-1,2-napthoquinone analog showed promising antimalarial activity in the bioassay. In contrast, a number of 2-hydroxy-1,4-naphthoquinones and dimeric quinones were less active.

Changes in cyclic nucleotide profiles in proliferating multi-cell spheroids.

The intracellular concentration of cyclic adenosine 3':5'-mono-phosphate (cAMP) has been shown to be related to each developmental phase of the cell cycle. Highest levels of this nucleotide are evident during the S-phase (the DNA synthetic phase) which has also been shown to be radiation-sensitive. The relationship between the levels of cyclic nucleotides, cAMP and guanosine 3':5'-monophosphate (cGMP), and the proliferation of cells in a tumor model system was investigated using V79-171b Chinese hamster lung cells grown both as monolayer and as three dimensional cell clusters (spheroids). The spheroid which is more radiation-resistant than its monolayer counterpart, has been used by many radiobiologists as an in vitro tumor model. Our results indicate that the yin-yang hypothesis of a opposing regulatory relation between the two different classes of cyclic nucleotides only held true for monolayer cultures (both exponential and plateau phase) but could not be demonstrated in the tumor model where the levels of both nucleotides increased directly with the diameter of the growing spheroid mass.